| 杜楠,裴雪涛,罗成基,李梁,冯凯,白慈贤,赵鹏.Egr-1启动子基因调控FLT3配基基因表达的实验研究[J].中华放射医学与防护杂志,2000,20(5):306-309 |
| Egr-1启动子基因调控FLT3配基基因表达的实验研究 |
| Studies on the expression of hematopoietic growth factors regulated by Egr-1 promoter |
| 投稿时间：1999-10-09 |
| DOI： |
| 中文关键词: 辐射 Egr1 启动子 FLT3配基 基因治疗 |
| 英文关键词:Radiation Egr-1 Promoter FLT3 ligand Gene therapy |
| 基金项目:国家自然科学基金资助项目(39900040);国家杰出青年自然科学基金资助项目(39825111) |
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| 摘要点击次数: 2658 |
| 全文下载次数: 1970 |
| 中文摘要: |
| 目的 探索辐射诱导基因调控元件启动的造血生长因子表达及其对造血的保护作用。方法 本实验将携带Egr1调控序列的FLT3配基(FL)和EGFP基因双顺反子载体(EgrEF)转染骨髓基质细胞系HFCL,采用流式细胞仪、RT-PCR、ELISA及细胞增殖法等观察细胞受照后Egr1调控元件诱导的FL表达及促进造血细胞的增殖作用。结果 在转染细胞HFCL/EF细胞中证实有外源性基因的整合和表达, 在25Gy辐射后16h的细胞培养上清液中表明FL含量较照射前明显增高((P<0.01),);辐射后10dHFCL/EF培养上清液对CD34+造血祖细胞的作用较辐射前具有明显的扩增作用((P<0.01),)。结论 在辐射后Egr1启动子调控的FL基因表达明显增高并具有保护造血作用。 |
| 英文摘要: |
| Objective To explore the regulatory effects of radiation-inducible gene on the expression of FLT3 ligand genes. Methods The human EL cDNA and EGFP cDNA were linked together with IRES and then inserted into the expression vector pCI-Egr-1, which was constructed by substituting CMV promoter in pCI-neo with the Egr-1 promoter(Egr-EF).The vector was transferred into human bone marrow stromal cell line HFCL by lipofectin TM .The cells were exposed to γ-radiation from 60 Co source at 0.5-20Gy.The contents of green fluorescence in HFCL/EF cells were detected with FACS.The expression of FL in HFCL/EF cells was confirmed with RT-PCR and ELISA respectively.The effects of FL in HFCL/EF cultural supernatants on expansion of CD34 + cells were also studied. Results The amounts of secreted FL in serum-free supernatants of Egr-EF were significantly higher than in the control group.EGFP and FL cDNA were successfully integrated and expressed in the cells,which were confirmed by FACS,RT-PCR and ELISA analysis.On day 10 of culture the number of CD34+ cells was significantly higher than that of the non-radiation group. Conclusion These in vitro data provide an experimental basis for the in vivo use of gene therapy of hematopoietic growth factor gene regulated by Egr-1 promoter to protect hematopoiesis from radiation injury. |
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